Fentanyl Transdermal Therapeutic System Provides Rapid Systemic Fentanyl

Absorption in Two Ball Pythons (Python regius)

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 Fentanyl Transdermal Therapeutic System
Provides Rapid Systemic Fentanyl Absorption in
Two Ball Pythons (Python regius)
Brett G. Darrow1, DVM, Gwen E. Myers2, DVM, Butch KuKanich3, DVM, PhD, DACVCP,
Kurt K. Sladky4,5, MS, DVM, DACZM, DECZM (Herpetology)
1. Department of Small Animal Surgery, College of Veterinary Medicine, University of Tennessee, Knoxville,
TN 37919, USA

2. Animal Health Services, Zoo Miami, Miami, FL 33177, USA

3. Department of Anatomy and Physiology, College of Veterinary Medicine, Kansas State University,
Manhattan, KS 66506, USA

4. Department of Surgical Sciences, School of Veterinary Medicine, University of Wisconsin, Madison, WI

5. Global Health Institute, School of Medicine and Public Health, University of Wisconsin, Madison, WI 53706,
USA

Abstract: Although our understanding of reptile analgesia continues to develop, µ-opioid receptor agonists are
currently the most effective analgesics in a variety of reptile species. Fentanyl, a potent, short-acting, µ- and
δ-opioid receptor agonist, is formulated for use as a transdermal therapeutic system (TTS) and was shown to be
absorbed across the skin of prehensile-tailed skinks (Corucia zebrata). Snakes, which possess highly permeable
skin, seem to be ideal candidates for effective transdermal analgesia. Fentanyl TTS (12 μg/h) was applied to the
midbody, dorsal surface of two ball pythons (Python regius). Specimen mean body weight was 2.60 kg (2.20–2.99
kg). Cardiac blood samples were collected for 7 days, and plasma fentanyl concentrations were determined using
high-performance liquid chromatography–mass spectroscopy. Therapeutic concentrations, as defined in mammals
(1 ng/ml), were reached within 4 h of patch application, and they were sustained throughout the study. Plasma
fentanyl steady-state (8.68–10.3 ng/ml) and maximum (11.7–17.6 ng/ml) concentrations were substantially higher
than those demonstrated to attain analgesia in mammals, suggesting the potential to use smaller dosages. Patches
remained well adhered throughout the study. Snakes exhibited no adverse side effects during the experiment. Data
from this study suggest that the TTS delivers quantifiable fentanyl concentrations in ball python plasma and may
serve as a route for prolonged analgesia administration to snakes. Additional research is needed to further
determine treatment efficacy and safety and to optimize the dosing regimen.

Key Words: ball python, fentanyl, analgesia, transdermal, Python regius, opioid.

Introduction noxious thermal stimulus (Sladky et al., 2007, 2008) and con-
tributed to a return to normal behavior in red-eared slider turtles
Reptiles comprise a substantial percentage of companion ani- undergoing unilateral gonadectomy (Kinney et al., 2011). Con-
mals, as well as zoological and laboratory research collections. sistent with these results, morphine treatment increases tail
Despite this, there is a considerable disparity and lack of funda- flick latencies in anole lizards (Anolis carolinensis) (Mauk et
mental knowledge with respect to understanding nociception al., 1981) and hot-plate withdrawal latencies in crocodiles
and antinociception in reptile species (Read, 2004; Sladky et (Crocodylus niloticus) (Kanui and Hole, 1992). Given that mor-
al., 2007, 2008). Current pain control methods for use in rep- phine is primarily a µ-opioid receptor agonist, antinociception
tiles rely on a few µ-opioid receptor agonists, partial agonists, in reptiles seems to occur with µ-opioid receptor agonist activa-
and partial antagonists for antinociception (Sladky et al., 2007, tion.
2008). Despite the questionable efficacy of butorphanol, a Fentanyl is a µ-opioid agonist, with 75–100 times the potency
µ-opioid receptor antagonist and κ-opioid receptor agonist, it of morphine, and is formulated for use as oral, inhalant, intrave-
remains a commonly used analgesic medication in clinical rep- nous, and transdermal applications. In mammals, fentanyl
tilian practice, highlighting a disconnect in the fundamental transdermal therapeutic systems (TTSs) deliver long-acting,
reptilian analgesic knowledge base (Read, 2004; Sladky et al., opioid-mediated analgesia to postoperative patients or patients
2007, 2008). suffering from long-term chronic pain. The time-release TTS
Previously, our lab demonstrated that morphine has antinoci- allows for noninvasive steady-state–like administration of fen-
ceptive properties in red-eared slider turtles (Trachemys scripta tanyl over a relatively long period (Kyles et al., 1996; Jeal and
elegans) and bearded dragons (Pogona vitticeps) exposed to a Benfield, 1997; Egger et al., 1998; Carroll et al., 1999; Lee et

94 Journal of Herpetological Medicine and Surgery Volume 26, No. 3–4, 2016
al., 2000; Glerum et al., 2001; Gellasch et al., 2002; Davidson zen plasma samples were submitted for PFC analysis at the
et al., 2004; Gamble, 2008; Hoy and Keating, 2008; Minville et study conclusion.
al., 2008). In addition to its long-acting analgesic potential, Immediately after blood sample collection, the snakes were
applying the fentanyl TTS modality to reptiles offers the bene- returned to their individual cages. To minimize any variation in
fits of eliminating injection site reactions, reducing handling metabolism that could potentially alter PFC, the animals were
stress for both the animal and people involved with restraint not fed during the 7 day study. They were allowed intermittent
during repeated injections, and increasing the safety of those access to water, under supervision, to avoid soaking the patches
handling dangerous animals. and potentially altering absorption rates. Blood sample collec-
To date, there are no published studies evaluating pharmaco- tion, under sevoflurane anesthesia, was repeated at the follow-
kinetics or efficacy of the fentanyl TTS in snakes. One report of ing time points: 4 h (T4), 8 h (T8), 12 h (T12), 24 h (T24), 36 h
TTS was recently published in prehensile-tailed skinks (Coru- (T36), 48 h (T48), 60 h (T60), 72 h (T72), 84 h (T84), 96 h (T96), 120
cia zebrata) and demonstrated that quantifiable plasma fentanyl h (T120), 144 h (T144), and 168 h (T168). In total, 12.7 and 9.4 ml/
concentrations (PFCs), sufficient for mammalian analgesia, kg of blood were drawn from snake 1 and 2, respectively.
could be attained in scaled reptilian species (Gamble, 2008). Immediately before the study, three other ball pythons within
Snake skin is used as a relevant in vitro model in human medi- the zoo’s quarantine facility developed clinical respiratory signs
cine because it is highly permeable (Panchagnula et al., 1997; and were diagnosed with bacterial pneumonia. No testing for a
Pongjanyakul et al., 2000). In fact, snake skin had the highest viral etiology was performed. Although both the zoo quarantine
diffusion rates of both polar and nonpolar compounds com- and study protocols required that the snakes in the study be
pared with 15 other types of animal skin (Panchagnula et al., maintained in isolated cages and gloves be worn and changed
1997). Therefore, snakes should make ideal reptile candidates with each handling, additional precautions were taken. Despite
to potentially benefit from TTS drug administration. We the possible complicating factors of medication coadministra-
hypothesized that the administration of the fentanyl TTS would tion, antibiotics were given to one snake at T144 for potential
result in transdermal diffusion of fentanyl with quantifiable respiratory difficulty. As this snake did not demonstrate further
concentrations in ball python (Python regius) plasma. respiratory compromise, its data were included in this study.
Medications administered to one snake included ceftazidime
Materials and Methods (Fortaz®, 20 mg/kg IM once, GlaxoSmithKline, Research Tri-
angle Park, NC) and enrofloxacin (Baytril®, 10 mg/kg IM
This project was approved by the Columbus Zoo and Aquar- once, Bayer Health Care LLC, Shawnee Mission, KS). Fluid
ium Conservation and Collection Management Committee. administration administered to both snakes consisted of lac-
Two adult, captive bred ball pythons of unknown sex were tated Ringer’s solution (5–7 ml/kg SC at T4, T12, T36, and T144 h;
quarantined and acclimated for 3 months at the Columbus Zoo Hospira Inc., Lake Forest, IL). All medications and fluids were
and Aquarium before beginning this study. The mean body administered in locations at least 20 cm from where the patch
weight was 2.60 kg (2.20–2.99 kg), with 3/5 body condition was placed.
scores. The snakes were monitored closely before, during, and The PFCs were quantified using previously published meth-
after the study. Each snake received a thorough physical exam- ods consisting of liquid chromatography (LC) (Shimadzu
ination, including a complete blood count and serum biochem- Prominence, Shimadzu Scientific Instruments, Columbia, MD)
istry screening, upon acquisition. Results were normal. Snakes and mass spectrometry (MS) (API 2000, Applied Biosystems,
were housed individually and maintained in a 12:12 h light:dark Foster City, CA) (Kukanich and Hubin, 2009). Fentanyl (Ceril-
cycle, ambient temperature range of 27.5–29.5°C (80–85°F), liant, Round Rock, TX), m/z 337→188, was quantified with
and humidity range of 35–50%. Each snake was fed appropri- fentanyl d5 (Cerilliant), m/z 341.4→188, as the internal stan-
ately sized killed mice at weekly intervals up to 2 days before, dard (IS) with the plasma standard curve linear from 0.5 to 500
and immediately after, the experiment. Individual microchips ng/ml. Plasma (0.5 ml) was added to 0.1 ml of IS, 0.5 ml 0.1 N
were placed subcutaneously for identification. sodium hydroxide, and vortexed for 5 sec. Drug was extracted
At the time of fentanyl TTS application, defined as T0, snakes from plasma using a solid phase extraction cartridge ([SPE],
were placed in a small Plexiglas chamber and briefly anesthe- Varian Bond Elut C18, Varian Inc., Palo Alto, CA). The SPEs
tized with sevoflurane gas for 5 min. The skin of each snake were conditioned with 1 ml of methanol, followed by 1 ml of
was cleaned of debris, if present, by using a dry cloth, but no deionized water. The plasma mixture was loaded, the SPEs
water or disinfectant was applied to avoid influencing transder- were rinsed with 1 ml of 5% methanol, and the drug was eluted
mal fentanyl absorption rates (U.S. Food and Drug Administra- with 1 ml of methanol. The eluate was evaporated to dryness at
tion, 2014). One fentanyl TTS (12.5 μg/h, Mylan Pharmaceuti- 40°C (104°F) under an airstream; reconstituted with 200 μl of
cals Inc., Morgantown, WV) was applied to the dorsum at 50% methanol; vortexed for 5 sec; and centrifuged for 5 min at
approximately the midbody and secured with four skin staples. 15,000 × g in a microcentrifuge tube to sediment particulates.
Staples were used to ensure that the patch would remain adhered The supernatant was transferred to an injection vial, and 50 μl
during the entire study. For blood sample collection, cardiocen- was injected into the LC/MS. The daily runs were accepted if a
tesis was necessary, because adequate quantities of blood are standard curve was linear, with a correlation coefficient (r)
difficult to collect using the ventral coccygeal vein in ball greater than 0.99, and if the measured standards were within
pythons. The ventral skin overlying the heart was disinfected 15% of the actual concentration. The accuracy of the assay was
with a 70% isopropyl soaked gauze pad, and approximately 1 102 ± 8% and the coefficient of variation was 6%.
ml of blood was collected for fentanyl analysis using a prehep- Pharmacokinetic analysis was performed with computer soft-
arinized, 25 gauge butterfly catheter. Blood was placed in a lith- ware (WinNonlin® 5.2, Pharsight Corporation, Mountain View,
ium heparin vial, and samples were immediately centrifuged at CA) by using noncompartmental methods. The maximum
3000 rpm for 16 minutes. The plasma supernatant was pipetted plasma concentration (CMAX) and time to CMAX (TMAX) were
into plasma banking vials and stored at −80°C (−112°F). Fro- determined directly from the data. The area under the plasma

Volume 26, No. 3–4, 2016 Journal of Herpetological Medicine and Surgery 95
 Figure 1. Individual time–plasma fentanyl concentration curves (overlaid) of ball pythons (N = 2) throughout 168 h application of a 12.5
μg/h patch. Subcutaneous fluids, marked by “F” on the x-axis, were administered at times T4, 12, 36, and 144. Antibiotics were administered as
indicated directly on the figure. E = enrofloxacin, C = ceftazidime.

concentration curve from T0 to the last measured time point mean CMAX of 14.7 ng/ml (11.7–17.6 ng/m). The OS was 21 and
(AUC0-LAST) was calculated using the linear trapezoidal method. 0% for snake 1 and 2, respectively. TMAX was at approximately
A steady-state concentration was defined as the average PFC 12 h and corresponded with the OS for the smaller snake 1,
from T24 to the last measured time point (CAVG 24-LAST). The over- whereas the larger snake 2 did not demonstrate an obvious
shoot (OS), expressed as a percentage, was defined as the dif- CMAX, TMAX, or OS. The mean CAVG 24-LAST was 9.50 ng/ml (8.68–
ference between the initial CMAX (before T24, if present) and 10.3 ng/ml). The TSS was 24 and 8 h for snake 1 and 2, respec-
CAVG 24-LAST expressed as a fraction of the CAVG 24-LAST or, OS = tively. Once steady state was achieved, it was maintained
100 × (CMAX − CAVG 24-LAST)/CAVG 24-LAST. The time to reach steady throughout the entire 7 day study, with minimal fluctuation in
state (TSS) was defined as the first time point after the initial OS PFC in both snakes. In general, there was a consistent PFC over
(if present) where the difference between the concentration at time profile produced by both snakes, with mild individual
the given time point and the CAVG 24-LAST is less than 50% of the fluctuation, as documented by the absorption curve profiles and
CAVG 24-LAST or, the time point at which (PFC − CAVG 24-LAST) < 0.5 pharmacokinetic parameters (Table 1).
(CAVG 24-LAST). The theoretical dose rate (micrograms per hour Throughout the study, the snakes did not seem to display
per kilogram) was estimated as the labelled fentanyl delivery obvious alterations in their normal behavior. The patch remained
from the TTS, 12 μ/h, divided by the animal’s weight (in kilo- well adhered to the skin of the snakes. Upon conclusion of the
grams). Mean theoretical dose of a group of animals was esti- study, the patches were removed and no gross skin inflamma-
mated as the labeled fentanyl delivery divided by a group’s tion was apparent. The two snakes began eating within 14 days
mean weight. The clearance per fraction of the dose absorbed of patch removal.
was calculated by dividing the dose rate by the CAVG 24-LAST.
Discussion
Results
Fentanyl TTS application administered to ball pythons pro-
Fentanyl was quantifiable in plasma, at concentrations vided higher plasma concentrations of fentanyl compared to
reported to be analgesic in humans and small mammalian spe- previous results reported in mammalian (Lee et al., 2000) and
cies (1 ng/ml), in both snakes within 4 h after fentanyl TTS skink (Gamble, 2008) studies, despite similar dosages (mean
administration and exceeded this concentration for the duration theoretical dose rates of ball pythons, 5.0 μg/kg/h; cats, 6.6 μg/
of the study (Fig. 1). Rapid fentanyl absorption resulted in a kg/h; and skinks, 2.6 μg/kg/h). The onset to detectable plasma

Table 1. Pharmacokinetic parameters of transdermal fentanyl in ball pythons (N = 2) after application of a 12.5 μg/h patch for 168 h.

AUC0-LAST CMAX TMAX OS CAVG 24-LAST TSS Theoretical dose rate CL/F

Parameter hr×ng/ml ng/ml h % ng/ml h μg/kg/h ml/kg/min

Snake 1 (2.20 kg) 1,790 17.6 11.9 21 10.3 24 5.7 9.2

Snake 2 (2.99 kg) 1,350 11.7 83.7 NA 8.68 8 4.2 8.1

Geometric mean 1,570 14.7 47.8 10.5 9.50 16 5.0 8.6

AUC0-LAST = area under the plasma concentration curve from T0 to the last measured time point, CMAX = maximum plasma concentration,
TMAX = time to CMAX, OS = overshoot, CAVG 24-LAST = steady-state concentration was defined as the average plasma fentanyl concentration
from time point 24 h to the last measured time point, TSS = time to reach steady state, CL/F = clearance per fraction of the dose absorbed,
NA = not applicable.

96 Journal of Herpetological Medicine and Surgery Volume 26, No. 3–4, 2016
concentration was ≤4 h. In other species, PFCs associated with actual dose rate could have been higher than 5.0 μg/kg/h. There-
analgesia include 0.2–2.0 ng/ml for humans (Kyles et al., 1996; fore, we cannot rule out the possibility that the actual plasma
Foley et al., 2001; Gamble, 2008), 1.56–1.73 ng/ml for cats clearance is higher than estimated clearance as we do not know
(Hofmeister and Egger, 2004), and as low as 0.6 ng/ml in dogs the actual dose rate. If a lower plasma clearance is truly present
(Hofmeister and Egger, 2004). PFCs ≥ 1 ng/ml were achieved in snakes, it may be due to slower fentanyl metabolism, as has
within ≤4 h of fentanyl patch placement in both snakes and been demonstrated in snakes, particularly in boid species
were maintained for the 7 day duration of study. The rate to (Andrews and Pough, 1985; Chappell and Ellis, 1987). Further
achieve ≥1 ng/ml in these snakes was more rapid than previ- studies could assess IV fentanyl to determine the actual clear-
ously reported in other species, including 24–36 h in prehensile ance of fentanyl in ball pythons.
tailed skinks (Gamble, 2008), 6–18 h in cats (Lee et al., 2000; Despite high CAVG 24-LAST, the duration of steady state was
Hofmeister and Egger, 2004), 12–24 h in dogs (Kyles et al., 144–160 h, which was the entire length of the study and may
1996; Egger et al., 1998; Hofmeister and Egger, 2004), and rab- also be consistent with a low systemic clearance. These concen-
bits (Foley et al., 2001). trations may continue beyond 7 days, but the TTS was removed
The fentanyl TTS in these snakes produced a substantially and no further sampling was performed. The duration of steady
higher mean CMAX of 14.7 ng/ml (11.7–17.6 ng/ml) and sus- state was much longer than is published in cats (up to 88 h, Lee
tained a higher mean CAVG 24-LAST of 9.50 ng/ml (8.68–10.3 ng/ et al., 2000; 65 h, Hofmeister and Egger, 2004), dogs (48 h,
ml) than has been reported in mammals or skinks. One snake Hofmeister and Egger, 2004), rabbits (up to 72 h, Foley et al.,
displayed an evident TMAX at T12, whereas the second snake did 2001), and skinks (80 h, Gamble, 2008).
not seem to have an obvious CMAX/TMAX. Cats receiving a High inter- and intraspecies variability have been reported
slightly higher dosage of transdermal fentanyl than these snakes using the fentanyl TTS (Carroll et al., 1999; Foley et al., 2001;
(cats, 6.6 μg/kg/h; snakes, 5.0 μg/kg/h) produced a mean CMAX Hofmeister and Egger, 2004; Gamble, 2008). Additional sam-
of 7 ng/ml, TMAX of 36 h, and steady-state PFC of 1.58 ng/ml, pling and a longer study period may be useful in establishing
and analgesic levels were achieved in only four (67%) of six of the duration of steady state in this species.
the cats after 12 h of patch application (Lee et al., 2000). The Because the TTS provides transdermal drug delivery, vari-
PFC observed in these snakes is also substantially higher than ability within integument will affect rates of fentanyl absorp-
that reported in prehensile-tailed skinks receiving 2.6 μg/kg/h. tion. Fick’s first law of diffusion implies that absorption is dic-
In skinks, the mean CMAX was 1.55 ng/ml at a mean TMAX of 67.6 tated by surface area (consistent with patch size), skin thickness,
h, whereas average skink steady-state levels were also much permeability, solubility within the skin, and temperature (affect-
lower than those of the snakes in this study (Gamble, 2008). ing blood flow and thus concentration gradient). Despite having
The TMAX of 12 h in one snake is consistent with what has been a similarly arranged dermis and epidermis, mammals and rep-
reported in cats (Hofmeister and Egger, 2004), dogs (Kyles et tiles differ such that mammals have hair (Gamble, 2008). Even
al., 1996), rabbits (Greenacre et al., 2005), and goats (Carroll et after shaving, minute hair shaft remnants may alter patch sur-
al., 1999). The rapid absorption and high CMAX in snakes are face contact and thus decrease fentanyl absorption (Foley et al.,
consistent with previous studies indicating that snakes possess 2001). This may partially explain the large discrepancy between
highly permeable skin with the capacity to quickly absorb a PFC achieved in cats and snakes, despite slightly higher dos-
wide range of polar and nonpolar, lipid and water-soluble com- ages used in cats (Lee et al., 2000). Even within reptile species,
pounds (Panchagnula et al., 1997; Pongjanyakul et al., 2000). the integument pattern seems to alter transdermal fentanyl
A steady-state condition was achieved by both snakes at a absorption (Gamble, 2008). An interlocking scale pattern that
mean TSS of 16 h (8–24 h). The fact that CAVG 24-LAST was much permits good surface contact, such as that of the prehensile
higher in these snakes than was previously reported in other tailed skink, is conducive to fentanyl transdermal absorption,
species, despite similar dosages, is likely the result of a combi- whereas a nodular pattern with limited surface contact, such as
nation of both effective transdermal absorption and lower rate that of the green iguana (Iguana iguana), is not as conducive to
of drug clearance. The actual clearance was not determined in transdermal fentanyl absorption (Gamble, 2008). The patch
this study, because we did not administer fentanyl IV. However, maintained good contact in both snakes throughout the duration
the clearance was estimated using the labeled drug delivery rate of this study. However, one post hoc conclusion was that skin
(12 μg/h in this study). With a mean of 8.6 ml/kg/min (8.1–9.2 staples were not necessary to maintain adherence to the skin,
ml/kg/min), these ball pythons have a much lower estimated and this has been documented in our laboratory multiple times
clearance fraction than is calculated in other animals. Using since this original study. In fact, staples could permit increased
mean weights, mean steady-state fentanyl concentrations, and fentanyl absorption either through primary disruption of the
patch sizes (including specified delivery rates) from species integument barrier or secondarily by inciting subclinical inflam-
analyzed in other studies, we noted that the mean clearance val- mation and altered perfusion to the area. Thus, we do not rec-
ues of other species were much higher: 56 ml/kg/min in skinks ommend the use of staples to secure the TTS in snakes. Stage of
(Gamble, 2008), 120 ml/kg/min in cats (Lee et al., 2000), and ecdysis also likely affects absorption of TTS medications. With
45–70 ml/kg/min in dogs (Egger et al., 1998). These calcula- recent ecdysis, diffusion thickness and diffusion barrier layers
tions, however, are based upon the package-stated fentanyl TTS are reduced because the integument no longer contains two lay-
dose delivery rate. In reality, the fentanyl TTS, designed for ers of epidermis separated by lymphatic fluid. This is consistent
human use, provides 19–59% of the available dose in cats and with previous published reports that snakes are more sensitive
27–99.6% in dogs (Lee et al., 2000). If the estimated plasma to trauma and toxins directly after ecdysis (Boyer and Boyer,
clearance of these ball pythons is the actual plasma clearance, it 1991; Cooper, 2006). Finally, subcutaneous fat is thought to act
is expected that the half-life of fentanyl in snakes would be lon- as a reservoir and creates a depot effect as has been described
ger than those reported in skinks (16 h; Gamble, 2008), cats (<6 previously (Lee et al., 2000; Gamble, 2008) and may retain
h; Lee et al., 2000), and dogs (<4 h; Egger et al., 1998). How- absorbed fentanyl from quickly entering systemic circulation.
ever, due to the likely increased permeability of snake skin, the In prehensile-tailed skinks, PFCs were maintained throughout a

Volume 26, No. 3–4, 2016 Journal of Herpetological Medicine and Surgery 97
3 day study and even increased in one individual directly after ing. Once analgesic levels are determined, it is possible lower
patch removal, likely as the result of deposition of lipid soluble dosages of the fentanyl TTS may be used.
fentanyl into subcutaneous adipose stores (Gamble, 2008). Dif- In two ball pythons, a single application of the fentanyl TTS
ferences in body weight, subcutaneous fat stores, skin thickness provided rapid absorption of fentanyl into the systemic circula-
(particularly the stratum corneum, Lee et al., 2000) as it relates tion, reaching >1 ng/m concentrations within 4 h of application.
to ecdysis, and the exact placement of the patch and staples, The PFCs achieved steady state and were maintained for 7
may account for the variation in PFC achieved by the fentanyl days. These results warrant future studies using larger snake
TTS observed in these snakes. However, drawing a definitive populations. Such studies should focus on determining analge-
conclusion from two individuals is difficult, and future studies sic efficacy, deleterious behavioral and physiological side
with larger numbers are warranted. effects, the development of scaled dosing regimens as were suc-
Non-integumentary related factors have been shown to influ- cessfully performed in small mammals (Lee et al., 2000; David-
ence fentanyl absorption and PFC. These factors may include son et al., 2004) and skinks by using metabolic energy formulae
environmental temperature and humidity, patch position and (Gamble, 2008), and the effects of shedding on transdermal
location, animal movement as it relates to TTS surface contact, drug absorption. Because reptile analgesia seems to be μ-opioid
cardiovascular and perfusion status, metabolic status, and indi- receptor mediated (Sladky et al., 2007, 2008), this study demon-
vidual patch variation (Riviere and Papich, 2001; Ashburn et strates that the fentanyl TTS has excellent potential as a valu-
al., 2003; Mills et al., 2004; Gamble, 2008). Many of these fac- able pain management tool in snakes.
tors were controlled during this study. Ambient temperature and
humidity were kept relatively constant throughout the study. Acknowledgments: This work was supported by the Colum-
Snakes were offered water, but not in a container large enough bus Zoo and Partners in Conservation Fund and University of
to submerse themselves, and only under supervision, thereby Wisconsin Merck-Merial Summer Scholars Research Program.
ensuring that the patch remained dry. The patch was placed
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